Nonsense Mutations in the Yeast SUP35 Gene Affect the [PSI+] Prion Propagation.
The essential SUP35 gene encodes yeast translation termination factor eRF3. Previously, we isolated nonsense mutations sup35-n and proposed that the viability of such mutants can be explained by readthrough of the premature stop codon. Such mutations, as well as the prion [PSI+], can appear in natural yeast populations, and their combinations may have different effects on the cells. Here, we analyze the effects of the compatibility of sup35-n mutations with the [PSI+] prion in haploid and diploid cells. We demonstrated that sup35-n mutations are incompatible with the [PSI+] prion, leading to lethality of sup35-n [PSI+] haploid cells. In diploid cells the compatibility of [PSI+] with sup35-n depends on how the corresponding diploid was obtained. Nonsense mutations sup35-21, sup35-74, and sup35-218 are compatible with the [PSI+] prion in diploid strains, but affect [PSI+] properties and lead to the formation of new prion variant. The only mutation that could replace the SUP35 wild-type allele in both haploid and diploid [PSI+] strains, sup35-240, led to the prion loss. Possibly, short Sup351-55 protein, produced from the sup35-240 allele, is included in Sup35 aggregates and destabilize them. Alternatively, single molecules of Sup351-55 can stick to aggregate ends, and thus interrupt the fibril growth. Thus, we can conclude that sup35-240 mutation prevents [PSI+] propagation and can be considered as a new pnm mutation.


Multidrug resistance ABC transporter structure predictions by homology modeling approaches.
Human multidrug resistance ABC transporters are ubiquitous membrane proteins responsible for the efflux of multiple, endogenous or exogenous, compounds out of the cells, and therefore they are involved in multi-drug resistance phenotype (MDR). They thus deeply impact the pharmacokinetic parameters and toxicity properties of drugs. A great pressure to develop inhibitors of these pumps is carried out, by either ligand-based drug design or (more ideally) structure-based drug design. In that goal, many biochemical studies have been carried out to characterize their transport functions, and many efforts have been spent to get high-resolution structures. Currently, beside the 3D-structures of bacterial ABC transporters Sav1866 and MsbA, only the mouse ABCB1 complete structure has been published at high-resolution, illustrating the tremendous difficulty in getting such information, taking into account that the human genome accounts for 48 ABC transporters encoding genes. Homology modeling is consequently a reasonable approach to overcome this obstacle. The present review describes, in the first part, the different approaches which have been published to set up human ABC pump 3D-homology models allowing the localization of binding sites for drug candidates, and the identification of critical residues therein. In a second part, the review proposes a more accurate strategy and practical keys to use such biological tools for initiating structure-based drug design.


A Stem-Loop Structure in Potato Leafroll Virus Open Reading Frame 5 (ORF5) Is Essential for Readthrough Translation of the Coat Protein ORF Stop Codon 700 Bases Upstream.
Translational readthrough of the stop codon of the capsid protein (CP) open reading frame (ORF) is used by members of the Luteoviridae to produce their minor capsid protein as a readthrough protein (RTP). The elements regulating RTP expression are not well understood, but they involve long-distance interactions between RNA domains. Using high-resolution mass spectrometry, glutamine and tyrosine were identified as the primary amino acids inserted at the stop codon of Potato leafroll virus (PLRV) CP ORF. We characterized the contributions of a cytidine-rich domain immediately downstream and a branched stem-loop structure 600 to 700 nucleotides downstream of the CP stop codon. Mutations predicted to disrupt and restore the base of the distal stem-loop structure prevented and restored stop codon readthrough. Motifs in the downstream readthrough element (DRTE) are predicted to base pair to a site within 27 nucleotides (nt) of the CP ORF stop codon. Consistent with a requirement for this base pairing, the DRTE of Cereal yellow dwarf virus was not compatible with the stop codon-proximal element of PLRV in facilitating readthrough. Moreover, deletion of the complementary tract of bases from the stop codon-proximal region or the DRTE of PLRV prevented readthrough. In contrast, the distance and sequence composition between the two domains was flexible. Mutants deficient in RTP translation moved long distances in plants, but fewer infection foci developed in systemically infected leaves. Selective 2'-hydroxyl acylation and primer extension (SHAPE) probing to determine the secondary structure of the mutant DRTEs revealed that the functional mutants were more likely to have bases accessible for long-distance base pairing than the nonfunctional mutants. This study reveals a heretofore unknown combination of RNA structure and sequence that reduces stop codon efficiency, allowing translation of a key viral protein.IMPORTANCE Programmed stop codon readthrough is used by many animal and plant viruses to produce key viral proteins. Moreover, such "leaky" stop codons are used in host mRNAs or can arise from mutations that cause genetic disease. Thus, it is important to understand the mechanism(s) of stop codon readthrough. Here, we shed light on the mechanism of readthrough of the stop codon of the coat protein ORFs of viruses in the Luteoviridae by identifying the amino acids inserted at the stop codon and RNA structures that facilitate this "leakiness" of the stop codon. Members of the Luteoviridae encode a C-terminal extension to the capsid protein known as the readthrough protein (RTP). We characterized two RNA domains in Potato leafroll virus (PLRV), located 600 to 700 nucleotides apart, that are essential for efficient RTP translation. We further determined that the PLRV readthrough process involves both local structures and long-range RNA-RNA interactions. Genetic manipulation of the RNA structure altered the ability of PLRV to translate RTP and systemically infect the plant. This demonstrates that plant virus RNA contains multiple layers of information beyond the primary sequence and extends our understanding of stop codon readthrough. Strategic targets that can be exploited to disrupt the virus life cycle and reduce its ability to move within and between plant hosts were revealed.


Au-Cu-M (M = Pt, Pd, Ag) nanorods with enhanced catalytic efficiency by galvanic replacement reaction.
This work reports a general wet-chemistry method to produce Au-Cu-X (X = Pt, Pd, and Ag) trimetallic nanorods using galvanic replacement reaction with Au-Cu nanorods as the templates. The mild conditions, such as low temperature and slow injection of metal precursors, contributed to the slow galvanic replacement reaction and helped keep the rod structure intact. The distribution of Au, Cu and the doping metals was even in the rods as confirmed by elemental mapping. The alloyed trimetallic nanorods showed enhanced catalytic activity for p-nitrophenol reduction after incorporating the third metal. Remarkably, the Au-Cu-Pd and Au-Cu-Pt nanorods show more than an order of magnitude improvement in the mass activities compared to the Au-Cu nanorods. This facile and general synthetic method can be applied to fabricate other multimetallic nanoparticles with varying shapes and compositions.


Cytologic characteristics of meningeal carcinomatosis: increased diagnostic accuracy using carcinoembryonic antigen and epithelial membrane antigen immunocytochemistry.
Traditionally, the diagnosis of meningeal carcinomatosis has been based on clinical suspicion and confirmed by cytologic study of cerebrospinal fluid. However, routine cytologic study may fail to detect malignant cells in a relatively large number of cases. We used immunocytochemistry in an attempt to increase the sensitivity of cytologic detection of malignant neoplasms in cerebrospinal fluid. Thirty-eight consecutive cerebrospinal fluid specimens from patients with clinically suspected meningeal carcinomatosis were selected for this study. Immunocytochemistry for carcinoembryonic antigen and epithelial membrane antigen were used on the archival Papanicolaou-stained cerebrospinal fluid preparations. Of the 23 specimens from patients with proven meningeal carcinomatosis, 13 were correctly diagnosed using cytomorphologic criteria alone. The diagnosis of malignant neoplasm in 8 cytologically suspicious and 1 cytologically negative specimen was confirmed using immunocytochemistry. All cases that were negative on follow-up were also negative cytologically and immunocytochemically. We conclude that in using common antibodies, such as carcinoembryonic antigen and epithelial membrane antigen, the sensitivity of the cytologic diagnosis of meningeal carcinomatosis increases, and that previously Papanicolaou-stained preparations are suitable for immunocytochemical studies.


Distinct mechanisms of forskolin-stimulated cyclic AMP accumulation and forskolin-potentiated hormone responses in C6-2B cells.
Forskolin activates a variety of adenylate cyclase systems and acts synergistically with receptor-mediated agonists which stimulate cyclic AMP production. The mechanism(s) and site(s) of forskolin action remain unclear. In C6-2B rat astrocytoma cells, forskolin stimulated greater than a 100-fold increase in cellular cyclic AMP content with a half-maximally effective concentration (EC50) of greater than 50 microM. Incubation of C6-2B cells with forskolin plus (-)-isoproterenol resulted in an increase in (-)-isoproterenol efficacy and potency. The EC50 for the forskolin-induced increase in (-)-isoproterenol potency was 22 nM, greater than 3 orders of magnitude lower than the EC50 for direct forskolin-stimulated cyclic AMP accumulation. Forskolin had no effect on beta-receptor affinity for (-)-isoproterenol as measured by competition for (-)-[125I]iodopindolol binding sites. Forskolin also augmented the responses to prostaglandin E1 and cholera toxin. Inhibition of protein synthesis with cycloheximide markedly reduced forskolin-stimulated cyclic AMP accumulation with little or no effect on the responses to (-)-isoproterenol, prostaglandin E1, or cholera toxin. The ability of forskolin to act synergistically with these agents was unaffected by cycloheximide treatment. These observations are compatible with a two-site model of forskolin action in C6-2B cells: a low-affinity site which mediates the direct action of forskolin to increase cellular cyclic AMP accumulation and a high-affinity site which mediates the potentiative action of forskolin. The low-affinity forskolin site appears to reside on a protein which is closely associated with the catalytic adenylate cyclase moiety and has a relatively shorter half-life than other components of the cyclase system. The high-affinity site resides on a more stable component of the adenylate cyclase system. The synergistic action of forskolin may involve an enhancement of the interaction between the guanine nucleotide-binding regulatory component and the catalytic component of the adenylate cyclase complex.


Positioning sensory terminals in the olfactory lobe of Drosophila by Robo signaling.
Olfactory receptor neurons and the interneurons of the olfactory lobe are organized in distinct units called glomeruli. We have used expression patterns and genetic analysis to demonstrate that a combinatorial code of Roundabout (Robo) receptors act to position sensory terminals within the olfactory lobe. Groups of sensory neurons possess distinct blends of Robo and Robo3 and disruption of levels by loss-of-function or ectopic expression results in aberrant targeting. In the wild type, most of the neurons send collateral branches to the contralateral lobe. Our data suggests that guidance of axons across brain hemispheres is mediated by Slit-dependent Robo2 signaling. The location of sensory arbors at distinct positions within the lobe allows short-range interactions with projection neurons leading to formation of the glomeruli.


Evaluation of the self-heating tendency of vegetable oils by differential scanning calorimetry.
The evaluation of the self-heating propensity of a vegetable (or animal) oil may be of significant importance during the investigation of a fire. Unfortunately, iodine value and gas chromatographic-mass spectrometric analysis do not lead to meaningful results in this regard. To the contrary, differential scanning calorimetry (DSC), which does not measure the chemical composition of the oil, but rather its thermodynamic behavior, produces valuable results. After a thorough literature review on the autooxidation of vegetable oils, several oils with different self-heating tendencies were analyzed using a Mettler-Toledo differential scanning calorimeter DSC 25 between 40 degrees C and 500 degrees C. Analyses were carried out both under air and nitrogen atmosphere to identify the phenomena due to autooxidation reactions. Using DSC, it was possible to observe the induction period of the oil (when available), the three different exothermic events, and the autoignition temperature (relatively independent of the oil type).


Cytological and biochemical correlation of late X-chromosome replication and gene inactivation in the mule.
The correlation between late X-chromosome replication and the quantitation of different X-linked glucose-6-phosphate dehydrogenase electrophoretic forms was studied in a natural hybrid, the female mule. In all four animals examined, a significant deviation from the expected 1: 1 ratio of random X-chromosome inactivation was observed, with the donkey X-chromosome the more frequently late-replicating one. The relative amounts of horse and donkey enzymes activities in lysates of mule skin fibroblasts and in peripheral blood were in agreement with this finding: the donkey enzyme was the minor component. Although random expression of the enzymes from the two parent species was not observed, sampling, selection, or adaptation may actually be responsible for the apparent "preferential inactivation". These studies support the hypothesis that late DNA replication indicates genetic inactivation.


Possible regulation of the Salmonella typhimurium histidine operon by adenosine triphosphate phosphoribosyltransferase: large metabolic effects.
An effort to find growth conditions leading to conditional regulation of the histidine operon of Salmonella typhimurium by the allosteric first enzyme of the pathway, adenosine triphosphate phosphoribosyltransferase (EC 22.214.171.124), is reported. A strain deleting the enzyme, TR3343, behaved simply and predictably under all growth conditions, whereas histidine auxotrophs containing active enzyme behaved in complicated ways dependent upon the location of the histidine pathway lesion. hisE strains derepressed the operon only one-half as much as TR3343 when grown on limiting histidine and a poor carbon source, but they also grew more slowly, probably as a result of high N1-(5-phospho-beta-D-ribosyl)-adenosine triphosphate levels in the cell. hisC strains exhibited oscillatory growth behavior and oscillatory histidine operon expression when grown on intermediate concentrations of the histidine precursor histidinol. This behavior probably was caused by synergistic in-phase variations in the histidine, purine nucleotide, and ppGpp pools of the cell. All of the growth and histidine operon expression effects associated with the presence of adenosine triphosphate phosphoribosyltransferase could be assigned to metabolic perturbation of the cell caused by unregulated enzymatic activity.


The shiverer mutation affects the persistence of Theiler's virus in the central nervous system.
Theiler's virus persists in the white matter of the spinal cord of genetically susceptible mice and causes primary demyelination. The virus persists in macrophages/microglial cells, but also in oligodendrocytes, the myelin-forming cells. Susceptibility/resistance to this chronic infection has been mapped to several loci including one tentatively located in the telomeric region of chromosome 18, close to the myelin basic protein locus (Mbp locus). To determine if the MBP gene influences viral persistence, we inoculated C3H mice bearing the shiverer mutation, a 20-kb deletion in the gene. Whereas control C3H mice were of intermediate susceptibility, C3H mice heterozygous for the mutation were very susceptible, and those homozygous for the mutation were completely resistant. This resistance was not immune mediated. Furthermore, C3H/101H mice homozygous for a point mutation in the gene coding for the proteolipid protein of myelin, the rumpshaker mutation, were resistant. These results strongly support the view that oligodendrocytes are a necessary viral target for the establishment of a persistent infection by Theiler's virus.


Measurements of OC and EC in coarse particulate matter in the southeastern United States.
The organic carbon (OC) and elemental carbon (EC) content of filter-based, 24-hr integrated particulate matter with aerodynamic diameters between 2.5 and 10 microm (PM10-2.5) was measured at two urban and two rural locations in the southeastern United States. On average, total carbon (OC + EC) comprised approximately 30% of PM10-2.5 mass at these four sites. Carbonate carbon was measured on a subset of samples from three sites and was found to be undetectable at a rural site in central Alabama, less than 2% of PM10-2.5 at an urban site in Georgia, and less than 10% of PM10-2.5 at an urban-industrial site in Alabama. Manual scanning electron microscopy (SEM) and computer-controlled SEM (CCSEM) along with energy dispersive X-ray spectroscopy (EDS) were used to identify individual carbonaceous particles in a selected subset of samples collected at one rural site and one urban-industrial site in Alabama. CCSEM results showed that biological material (e.g., fungal spores, pollen, and vegetative detritus) accounted for 60-70% of the carbonaceous mass in PM10-2.5 samples with concentrations in the range of 2-16 microg/m3. Samples with higher PM10-2.5 concentrations (25-42 microg/m3) at the urban-industrial site were found by manual SEM to have significant amounts of unidentified carbonaceous material, likely originating from local industrial activities. Both filter-based OC and EC concentrations and SEM-identified biological material tended to have higher concentrations during warmer months. Upper limits for organic mass (OM) to OC ratios (OM/OC) are estimated for PM10-2.5 samples at 2.1 for urban sites and 2.6-2.7 for rural sites.


Development of peripheral autonomic synapses: neurotransmitter receptors, neuroeffector associations and neural influences.
1. The functional innervation of autonomic target tissues occurs early during development, at a time when both the nerves and post-synaptic target tissues are still differentiating. 2. Physiological responses appear soon after the arrival of the first fibres when uptake and release mechanisms within the nerves are already functional. Initial responses differ from those in the mature animal, both in the form and, frequently, in the subtypes of receptors involved. 3. Results of a number of studies suggest that the initial expression of neurotransmitter receptors during development is largely independent of neural influences. Changes recorded in neurotransmitter receptor expression during development appear to be similarly independent of neural influences. 4. While signal transduction pathways coupling adrenergic neurotransmitter receptors to effector responses appear to develop independently of the nerves, the efficient coupling of muscarinic receptors often requires the action of the neurotransmitter, acetylcholine. 5. During the period of synapse formation, the neural plexus continues to expand. While developing varicosities can release the neurotransmitter, the capacity for neurotransmitter retention appears to be restricted. Developmental changes in the neurotransmitters that produce functional responses, while well known in the sweat glands, may also be seen in more subtle forms in other target tissues. 6. Ultrastructural studies suggest that close physical associations between the membranes of the release sites of the developing nerves and the target cells may form early during development when physiological responses are still immature. These close associations could enable more specific reciprocal interactions between nerves and target cells involving known and novel growth factors, neuropeptides and cytokines important in shaping the mature synaptic characteristics.


Multimodal imaging in infectious and noninfectious intermediate, posterior and panuveitis.
Given the heterogeneity of uveitis, markers of inflammation vary from patient to patient. Multimodal imaging has proven itself to be critical for accurate evaluation for disease activity and treatment response in uveitis. Ultra-widefield (UWF) fluorescein angiography and autofluorescence (AF) as well as optical coherence tomography angiography (OCTA) have provided insights into disease pathogenesis and monitoring not previously appreciated. In addition to structural retinal imaging, OCT can be used to assess the choroid, the posterior cortical vitreous and the retinal vasculature in eyes with uveitis. Multimodal ocular imaging in eyes with uveitis is critical for disease diagnosis and assessing response to treatment. UWF fluorescein angiography can detect retinal vasculitis even in the absence of overt vascular sheathing. UWF AF can help detect more chorioretinal lesions than clinically visible. OCT can be used to assess the posterior cortical vitreous, retina, large retinal vessels and choroid in uveitis. The use of multimodal imaging will likely be needed to determine clinical trial endpoints in studies evaluating therapeutics for uveitis.


[Artificial circulation in high-risk percutaneous coronary interventions].
In their everyday practical clinical work cardiovascular surgeons sometimes have to deal with patients at extremely high risk of both percutaneous coronary interventions (PCIs) and direct myocardial revascularization. A method of choice in such situations may become a PCI supported by artificial circulation (AC), for which foreign and Russian authors propose using systems of prolonged extracorporeal membrane oxygenation (ECMO). The present work was aimed at sharing our experience with using standard systems of AC and their modifications (mini-circuit systems) for performing high-risk PCIs. Between October 2011 and November 2014, PCIs supported by artificial circulation were performed in a total of ten patients. All had extremely high risk of PCI due to coronary artery lesions [subocclusion of the trunk of the left coronary artery (LCA) combined with occlusion or significant stenosis of the right coronary artery (RCA)], concomitant pathology (obesity, diabetes mellitus, age, etc.) or critical state (circulatory arrest, resuscitating measures). Three patients during PCI developed ventricular fibrillation and one patient suffered an episode of asystole. All cardiac arrhythmias after restoration of the coronary blood flow disappeared spontaneously on the background of extracorporeal support. The only lethal outcome was registered during emergency PCI in a female patient admitted to the roentgen-operating room in the state of clinical death, on the background of continuing resuscitation measures. The presented methods of assisted circulation based on the standard AC systems and modification thereof (mini-circuit system) proved efficient. They make it possible to perform high-risk PCIs, including in clinics having neither appropriate equipment nor experience in ECMO.


CGP 41251 and tamoxifen selectively inhibit mitogen-activated protein kinase activation and c-Fos phosphoprotein induction by substance P in human astrocytoma cells.
The substance P (SP) receptor (NK-1 subtype) is widely expressed in primary human astrocytomas and glioblastomas and many brain tumor-derived cell lines. SP receptor activation stimulates the mitogen-activated protein (MAP) kinase pathway and the expression of immediate-early genes (e.g., c-Fos and c-Myc), resulting in an increase in DNA synthesis in human astrocytoma U-373 MG cells. In this study, we investigated the role of protein kinase C (PKC) in SP receptor activation of the MAP kinase pathway. SP peptide, epidermal growth factor, and the PKC activator 12-O-tetradecanoylphorbol-13-acetate (TPA) induced the tyrosine phosphorylation of the Erk1 and Erk2 MAP kinases in a concentration-dependent manner in U-373 MG cells. Pretreatment of the cells with PKC inhibitors, CGP 41251 or tamoxifen, inhibited tyrosine phosphorylation of Erk1 and Erk2 MAP kinases induced by low concentrations of SP or TPA and significantly attenuated phosphorylation at high concentrations of SP or TPA. The inhibitory effect exhibited by tamoxifen on SP-induced MAP kinase activation is similar to that exhibited by the selective PKC inhibitor CGP 41251, suggesting that the PKC enzyme is the in situ target for both inhibitors. Furthermore, SP-induced c-Fos phosphoprotein expression is inhibited by CGP 41251 or tamoxifen with similar efficacy. Importantly, neither CGP 41251 nor tamoxifen has any detectable effect on the MAP kinase activation by epidermal growth factor, consistent with the ability of this growth factor to activate the MAP kinase pathway by a PKC-independent mechanism. Prolonged treatment with TPA resulted in down-regulation of PKC and selective inhibition of TPA- and SP-induced Erk1 and Erk2 tyrosine phosphorylation in U-373 MG cells. Consistent with the in situ results, CGP 41251 and tamoxifen significantly inhibited endogenous PKC enzymatic activity from U-373 MG cells in vitro. In contrast to CGP 41251 and tamoxifen, Gö 6976, a highly selective inhibitor for PKC alpha and PKC beta 1 isozymes, did not inhibit SP- or TPA-induced tyrosine phosphorylation of Erk1 and Erk2 MAP kinases; rather, it inhibited a signaling pathway leading to the phosphorylation of cAMP-responsive element binding protein in U-373 MG cells. To investigate whether selective PKC isozyme(s) are involved in the activation of the MAP kinase pathway by SP, we determined the expression of PKC isozymes in U-373 MG cells. We found that U-373 MG cells express nine different PKC isozymes (alpha, beta I, beta II, epsilon, delta, eta, zeta, iota, and mu) and that stimulation with SP results in significant and selective translocation of PKC epsilon isozyme from cytosolic to membrane fraction. This establishes a correlation between the ability of SP to activate the MAP kinase pathway and its ability to translocate PKC epsilon. In conclusion, the results presented in this study demonstrate that SP receptor activation of PKC, possibly PKC epsilon, leads to the activation of the MAP kinase pathway, and that this pathway can be inhibited by known PKC inhibitors.


Response of the non-pregnant and pregnant rat uterus to chloroquine in vitro.
The contractility of the uterus is known to vary at different phases of the oestrus cycle and during pregnancy. Chloroquine in low doses produced an initial reduction of the amplitude of spontaneous contraction followed by a dose-dependent increase in the amplitude of contraction and reduction in the rate of contraction of uteri in the proestrus, oestrus and dioestrus phases of the oestrus cycle. Reduction in both the rate and amplitude of contraction was recorded to higher doses on the uterus in the proestrus phase whilst a further increase in amplitude and reduction in rate of contraction were observed on uteri in oestrus and dioestrus phases. Chloroquine produced irregular non-dose-dependent spontaneous contractions of the quiet uterus in early pregnancy.


Transplant immunosuppression enhances efficiency of adenoviral-mediated gene retransfection: inhibition of interferon-gamma and immunoglobin G.
Transplant immunosuppression regimen facilitates successful adenovirus-mediated gene transfection and retransfection in the rat lung. Herein, we investigated the effect of this strategy on circulating cytokines and antiadenoviral immunoglobin G antibody. Male Lewis rats were transfected with 1 x 10(9) pfu/mL of E1-deleted Ad5CMVLacZ vector transtracheally. Rats were randomly assigned to receive daily intraperitoneal triple immunosuppression regimen consisting of cyclosporine (15 mg/kg per day), azathioprine (6 mg/kg per day), and methylprednisolone (2.5 mg/kg per day), or normal saline solution. Retransfection was performed 35 days later to all nonimmunosuppressed animals, whereas immunosuppressed rats were further randomized to receive retransfection or phosphate-buffered saline. Animals were sacrificed on days 1, 2, 7, 35, 42, and 49 after the initial transfection. Beta-galactosidase activity was measured on lung homogenates. Interferon-gamma, tumor necrosis factor-alpha, and antiadenoviral immunoglobin G were measured from the serum. Enhanced and prolonged transgene expression was observed in immunosuppressed animals, especially after retransfection. Concentrations of serum tumor necrosis factor-alpha in both groups were less than 12 pg/mL throughout the study. A significant increase in serum interferon-gamma levels was observed in nonimmunosuppressed animals after retransfection; this was not seen in the immunosuppressed animals. Serum antiadenoviral immunoglobin G titers in both groups were sharply elevated on day 1, and declined to basal levels by day 7, reflecting a preexisting level of humoral immunity to adenovirus. The titer in nonimmunosuppressed rats was significantly increased after retransfection, but remained at very low level in immunosuppressed animals. Inhibition of interferon-gamma and antiadenoviral immunoglobin G production by triple immunosuppressants may be part of the mechanisms that lead to enhanced and prolonged transgene expression after retransfection.


Industrial research in america: challenge of a new synthesis.
Industrial research in America faces a new era. Success will depend on a synthesis utilizing the strengths of industry, government, and academia to achieve industrial innovation. The origins of U.S. industrial research in the 19th century have led to its current traditions and philosophy. Vigorous, diverse, animated by both basic scientific knowledge and empirical techniques, it is today channeled by both societal needs and new technical possibilities. Many of its efforts involve fundamental research. Meeting future expectations will require even broader science, much of it from academia, as well as a license to function from governments and society.


Benefit finding in survivors of childhood cancer and their parents: further empirical support for the Benefit Finding Scale for Children.
Diagnosis of a life-threatening condition has been linked to post-traumatic stress. However, only recently has it been acknowledged that positive outcomes including post-traumatic growth or benefit finding may also occur. The aim of our study was to extend previous work describing benefit finding among survivors of childhood cancer, by determining the contribution of demographic and medical variables and associations between child benefit finding and parent post-traumatic growth. Survivors of any child cancer (leukaemia, central nervous system or solid tumour; age 12-15 years; completed treatment >2 years) were recruited from routine follow-up clinics and asked to complete questionnaires [Benefit Finding Scale for Children (BFSC), quality of life (QOL), post-traumatic stress (PTS), illness perception and optimism]. Parents completed parallel measures to describe their own post-traumatic growth (PTG), QOL, PTS and illness perception. Forty-eight survivors and parents completed questionnaires (response rate: 81%). The BFSC showed good internal reliability (alpha = 0.91). Diagnosis of leukaemia, greater optimism and reports that the illness still affects their life today were associated with higher scores on the BFSC among survivors themselves. For parents, perceptions of how much the illness still affects them emotionally was associated with PTG. There was no association between children's benefit finding and parents' PTG. The BFSC is a useful and reliable instrument to assess positive outcomes after cancer in children. The extent to which survivors are optimistic and perceive on-going effects of the illness on their daily lives is significantly associated with the ability to find benefit after end of treatment.


Catastrophic wear in a metal-on-ceramic total hip arthroplasty.
A 51-year-old woman came to our clinic 6 months after a right total hip arthroplasty. She had noticed a slowly growing mass in the proximal thigh and referred progressive pain in the right groin. Plain radiography revealed premature acetabular cup aseptic loosening, and in the computed tomography study, a 14-cm-diameter mass was observed. Revision surgery was performed, showing a metal-on-ceramic bearing surface. The histologic analysis of surrounding tissues was reported as massive metallosis. Although occasionally chosen for primary or revision hip arthroplasty, there is little information available about the in vivo wear behavior of this combination. This important fact should be taken into account before considering such a surface alternative.


The Assessment of Preterm Infants' Behavior (APIB): furthering the understanding and measurement of neurodevelopmental competence in preterm and full-term infants.
The Assessment of Preterm Infants' Behavior (APIB) is a newborn neurobehavioral assessment appropriate for preterm, at risk, and full-term newborns, from birth to 1 month after expected due date. The APIB is based in ethological-evolutionary thought and focuses on the assessment of mutually interacting behavioral subsystems in simultaneous interaction with the environment. The subsystems of functioning assessed include the autonomic (respiration, digestion, color), motor (tone, movement, postures), state organization (range, robustness, transition patterns), attention (robustness, transitions), and self-regulation (effort, success) systems as well as the degree of facilitation required to support reorganization and subsystem balance. The environment is represented by a sequence of distal, proximal, tactile, and vestibular challenges, derived from the BNBAS. The APIB conceptualizes infant competence as the degree of differentiation of subsystem function and degree of modulation of subsystem balance at any stage in infant development. Infants are understood as actively seeking their next differentiation, while counting on good enough environments to assure progressing developmental competence. In the case of interference such as premature birth, the mismatch of expectation and actual experience causes misalignment, which may become developmentally costly. The assessment is a finely tuned dialogue between examiner and infant, which requires training, skill and self-knowledge. The APIB has well established inter-rater-reliability, concurrent and construct validity, and is clinically relevant for behavioral intervention and individually appropriate and supportive care.


New model of PIRADS and adjusted prostatespecific antigen density of peripheral zone improves the detection rate of initial prostate biopsy: a diagnostic study.
This study explored a new model of Prostate Imaging Reporting and Data System (PIRADS) and adjusted prostate-specific antigen density of peripheral zone (aPSADPZ) for predicting the occurrence of prostate cancer (PCa) and clinically significant prostate cancer (csPCa). The demographic and clinical characteristics of 853 patients were recorded. Prostate-specific antigen (PSA), PSA density (PSAD), PSAD of peripheral zone (PSADPZ), aPSADPZ, and peripheral zone volume ratio (PZ-ratio) were calculated and subjected to receiver operating characteristic (ROC) curve analysis. The calibration and discrimination abilities of new nomograms were verified with the calibration curve and area under the ROC curve (AUC). The clinical benefits of these models were evaluated by decision curve analysis and clinical impact curves. The AUCs of PSA, PSAD, PSADPZ, aPSADPZ, and PZ-ratio were 0.669, 0.762, 0.659, 0.812, and 0.748 for PCa diagnosis, while 0.713, 0.788, 0.694, 0.828, and 0.735 for csPCa diagnosis, respectively. All nomograms displayed higher net benefit and better overall calibration than the scenarios for predicting the occurrence of PCa or csPCa. The new model significantly improved the diagnostic accuracy of PCa (0.945 vs 0.830, P < 0.01) and csPCa (0.937 vs 0.845, P < 0.01) compared with the base model. In addition, the number of patients with PCa and csPCa predicted by the new model was in good agreement with the actual number of patients with PCa and csPCa in high-risk threshold. This study demonstrates that aPSADPZ has a higher predictive accuracy for PCa diagnosis than the conventional indicators. Combining aPSADPZ with PIRADS can improve PCa diagnosis and avoid unnecessary biopsies.


[Translated article] Hip arthroscopy for femoroacetabular impingement with 10-year minimum follow-up.
To determine outcomes after hip arthroscopy (HA) for femoroacetabular impingement (FAI) at a minimun 10-year follow up and identified risk factors for revision surgery. Retrospective study of a prospective database of patients undergoing HA between January 2010 and December 2011. Rim chondral injuries were evaluated using the acetabular labral articular disruptions system (ALAD). Clinical and radiological data was obtained. Cumulative survival was estimated by Kaplan-Meier and a multivariate Cox proportional hazards model. Seventy-four patients were included in the study. Mean follow-up was 132 months. There was statistically significant improvement from preoperative PROs at latest follow-up. Seventeen patients (23.9%) underwent revision surgery. Odds ratio for revision surgery was 8.08 in a patient above 40 years old, 7.57 in a patient Tönnis greater than 1, and 4.25 in a patient ALAD 2-3. Cumulative survivorship rate at 10 years was 77.8%, with a 45.4% for patients with Tönnis grade greater than 1 in front of 85.2% for patients with Tönnis grade of 1 or less (P<.001). Risk factor for revision surgery was preoperative degree of osteoarthrosis (P=.02). Cumulative survivorship rate at 10 years was 45.4% for patients with Tönnis grade greater than 1 in front of 85.2% for patients with Tönnis grade of 1 or less (P<.001). Age, chondral injuries, and degree of osteoarthrosis would increase the risk for revision surgery.


Genetic analysis of contributions of dorsal group and JAK-Stat92E pathway genes to larval hemocyte concentration and the egg encapsulation response in Drosophila.
Drosophila larvae defend themselves against parasitoid wasps by completely surrounding the egg with layers of specialized hemocytes called lamellocytes. Similar capsules of lamellocytes, called melanotic capsules, are also formed around "self" tissues in larvae carrying gain-of-function mutations in Toll and hopscotch. Constitutive differentiation of lamellocytes in larvae carrying these mutations is accompanied by high concentrations of plasmatocytes, the major hemocyte class in uninfected control larvae. The relative contributions of hemocyte concentration vs. lamellocyte differentiation to wasp egg encapsulation are not known. To address this question, we used Leptopilina boulardi to infect more than a dozen strains of host larvae harboring a wide range of hemocyte densities. We report a significant correlation between hemocyte concentration and encapsulation capacity among wild-type larvae and larvae heterozygous for mutations in the Hopscotch-Stat92E and Toll-Dorsal pathways. Larvae carrying loss-of-function mutations in Hopscotch, Stat92E, or dorsal group genes exhibit significant reduction in encapsulation capacity. Larvae carrying loss-of-function mutations in dorsal group genes (including Toll and tube) have reduced hemocyte concentrations, whereas larvae deficient in Hopscotch-Stat92E signaling do not. Surprisingly, unlike hopscotch mutants, Toll and tube mutants are not compromised in their ability to generate lamellocytes. Our results suggest that circulating hemocyte concentration and lamellocyte differentiation constitute two distinct physiological requirements of wasp egg encapsulation and Toll and Hopscotch proteins serve distinct roles in this process.


Body weight and tympanic temperature change in professional rugby league players during night and day games: a study in the field.
This study investigated the impact of day and night games in the professional rugby league on body weight and tympanic temperature change in participants. Twenty-five players contracted to an English Super League club had their pre- and postgame body weight and tympanic temperatures recorded during 10 games played during the official professional rugby league season, representing a total of 165 player appearances. The mean (+/-SD) ambient temperature and relative humidity was 12.3 degrees C (+/-6.0) and 83.3% (+/-11.4), respectively. Body weight was recorded using a set of calibrated Soehnle digital scales with players wearing underwear only and towel-dried of all sweat (postmatch). Tympanic temperature was recorded using a Braun ThermoScan Pro LT instant thermometer. Players were allowed to ingest fluid ad libitum throughout each match. Wet and dry bulb temperatures were recorded at the commencement and completion of each match. Significant changes in pregame to postgame body weight and tympanic temperature were found, but these were not influenced by the time of day that the game was played. The mean decrease in body weight was 0.86 kg (SE 0.085, p < 0.000), and the mean increase in tympanic temperature was 0.34 degrees C (SE 0.070, p < 0.000). No significant differences in body weight or tympanic temperature change were found between forwards and backs. Participation in the English professional rugby league can produce significant decreases in body weight and increases in body temperature that may lead to impaired performance. It is important for participants, coaches, and administrators to introduce strategies that will minimize the impact of environmental conditions on thermoregulation and ultimately player performance.


[Body constitution in children in Cuentepec, Morelos].
A somatometric estimation of the body constitution was made in 87 females and 72 males applying the values of weight and height, the muscle circumference and medial section of the left arm and fat area of that section. The material represented 28.57 and 23.61% respectively of the total number of the 5 to 19 years-old inhabitants of the Nahua community of Cuentepec, Morelos. Weight and height, -but specially the latter-were impaired, both in males as in females, but more markedly in the latter. The weight as expected for the height was not affected. The behavior of the total area of the arm was quite similar to that of the body weight. In contrast, in females under 14 years old, the muscle area of the arm was located as an average, slightly below the limits of normality; subsequently, it was normal. In males it was significantly lower than in females. The fat area was the most deteriorated and the defect was persistently greater in females than in males. All this means protection mechanisms of the active tissue mass (homeorrhesis) leading to a greater resistance in females, but at the same time, being responsible for a decrease in magnitude of the somatometric expression of the sexual dimorphism.


Retinal biomarkers and pharmacological targets for Hermansky-Pudlak syndrome 7.
Deletion of dystrobrevin binding protein 1 has been linked to Hermansky-Pudlak syndrome type 7 (HPS-7), a rare disease characterized by oculocutaneous albinism and retinal dysfunction. We studied dysbindin-1 null mutant mice (Dys-/-) to shed light on retinal neurodevelopment defects in HPS-7. We analyzed the expression of a focused set of miRNAs in retina of wild type (WT), Dys+/- and Dys-/- mice. We also investigated the retinal function of these mice through electroretinography (ERG). We found that miR-101-3p, miR-137, miR-186-5p, miR-326, miR-382-5p and miR-876-5p were up-regulated in Dys-/-mice retina. Dys-/- mice showed significant increased b-wave in ERG, compared to WT mice. Bioinformatic analysis highlighted that dysregulated miRNAs target synaptic plasticity and dopaminergic signaling pathways, affecting retinal functions of Dys-/- mice. Overall, the data indicate potential mechanisms in retinal neurodevelopment of Dys-/- mice, which may have translational significance in HSP-7 patients, both in terms of diagnostic/prognostic biomarkers and novel pharmacological targets.


Docosahexaenoic acid but not eicosapentaenoic acid lowers ambulatory blood pressure and heart rate in humans.
Animal studies suggest that the 2 major omega3 fatty acids found in fish, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), may have differential effects on blood pressure (BP) and heart rate (HR). The aim of this study was to determine whether there were significant differences in the effects of purified EPA or DHA on ambulatory BP and HR in humans. In a double-blind, placebo-controlled trial of parallel design, 59 overweight, mildly hyperlipidemic men were randomized to 4 g/d of purified EPA, DHA, or olive oil (placebo) capsules and continued their usual diets for 6 weeks. Fifty-six subjects completed the study. Only DHA reduced 24-hour and daytime (awake) ambulatory BP (P<0.05). Relative to the placebo group, 24-hour BP fell 5.8/3.3 (systolic/diastolic) mm Hg and daytime BP fell 3.5/2.0 mm Hg with DHA. DHA also significantly reduced 24-hour, daytime, and nighttime (asleep) ambulatory HRs (P=0. 001). Relative to the placebo group, DHA reduced 24-hour HR by 3. 5+/-0.8 bpm, daytime HR by 3.7+/-1.2 bpm, and nighttime HR by 2. 8+/-1.2. EPA had no significant effect on ambulatory BP or HR. Supplementation with EPA increased plasma phospholipid EPA from 1. 66+/-0.07% to 9.83+/-0.06% (P<0.0001) but did not change DHA levels. Purified DHA capsules increased plasma phospholipid DHA levels from 4.00+/-0.27% to 10.93+/-0.62% (P<0.0001) and led to a small, nonsignificant increase in EPA (1.52+/-0.12% to 2.26+/-0.16%). Purified DHA but not EPA reduced ambulatory BP and HR in mildly hyperlipidemic men. The results of this study suggest that DHA is the principal omega3 fatty acid in fish and fish oils that is responsible for their BP- and HR-lowering effects in humans. These results have important implications for human nutrition and the food industry.


Prognostic value of mid-regional pro-adrenomedullin levels taken on admission and discharge in non-ST-elevation myocardial infarction: the LAMP (Leicester Acute Myocardial Infarction Peptide) II study.
The purpose of this study was to assess the prognostic value of admission and discharge mid-regional pro-adrenomedullin (sAM) levels in non-ST-elevation myocardial infarction (MI) and identify values to aid clinical decision making. N-terminal pro-B-type natriuretic peptide and GRACE (Global Registry of Acute Coronary Events) score were used as comparators. sAM is a stable precursor of adrenomedullin. We measured plasma sAM on admission and discharge in 745 non-ST-elevation MI patients (514 men, median age 70.0 +/- 12.7 years). The primary end point was a composite of death, heart failure, hospitalization, and recurrent acute MI over mean follow-up of 760 days (range 150 to 2,837 days), with each event assessed individually as secondary end points. During follow-up, 120 (16.1%) patients died, and there were 65 (8.7%) hospitalizations for heart failure and 77 (10.3%) recurrent acute MIs. Both admission and discharge levels were increased (median 0.81 nmol/l [range 0.06 to 5.75 nmol/l] and 0.76 nmol/l [range 0.25 to 6.95 nmol/l], respectively) compared with established normal ranges. Multivariate adjusted Cox regression models revealed that both were associated with the primary end point (hazard ratio: 9.75 on admission and 7.54 on discharge; both p < 0.001). Admission sAM was particularly associated with early (<30 days) mortality (c-statistic = 0.90, p < 0.001), and when compared with N-terminal pro-B-type natriuretic peptide and GRACE score, it was the only independent predictor of this end point. Admission sAM >1.11 nmol/l identified those at highest risk of death (p < 0.001). Patients with above-median admission sAM may benefit from revascularization. sAM level is prognostic for death or heart failure. Admission levels are a strong predictor of early mortality and, when >1.11 nmol/l, complements the GRACE score to improve risk stratification.


Preventing Intra-hospital Infection and Transmission of Coronavirus Disease 2019 in Health-care Workers.
Coronavirus disease 2019 poses an occupational health risk to health-care workers. Several thousand health-care workers have already been infected, mainly in China. Preventing intra-hospital transmission of the communicable disease is therefore a priority. Based on the Systems Engineering Initiative for Patient Safety model, the strategies and measures to protect health-care workers in an acute tertiary hospital are described along the domains of work task, technologies and tools, work environmental factors, and organizational conditions. The principle of zero occupational infection remains an achievable goal that all health-care systems need to strive for in the face of a potential pandemic.


Cloprostenol injection improves reproductive characteristics in low libido Iranian Holstein bulls.
The objective of this study was to determine whether injections of Cloprostenol (PGF2alpha analog) in low libido Holstein bulls can improve seminal characteristics and libido. Ten low libido Iranian Holstein bulls were randomly assigned to two groups and received; (1) 250 microg ofCloprostenol (n=5) or (2) 2 mL of saline (n=5) 30 min prior to collection of semen 2 days per week for 2 months. Reaction time was significantly decreased in treatment group. Duration of ejaculation was significantly increased in treatment group. Semen volume and sperm concentration were greater in treated bulls in compare with controls. The percentage of morphologically normal sperm cells, percentage of live sperm cells, motile sperm cells and post-thaw motile sperm cells were not affected by treatment. Plasma testosterone concentrations were increased approximately two fold after Cloprostenol injection. Overall, injection of Cloprostenol at this dosage and frequency increased libido, semen volume, sperm concentration and plasma testosterone concentration in low libido Holstein bulls.


Intestinal flora study reveals the mechanism of Danggui Shaoyao San and its decomposed recipes to improve cognitive dysfunction in the rat model of Alzheimer's disease.
Alzheimer's disease (AD), characterized by a severe decline in cognitive function, significantly impacts patients' quality of life. Traditional Chinese Medicine (TCM) presents notable advantages in AD treatment, closely linked to its regulation of intestinal flora. Nevertheless, a comprehensive exploration of the precise role of intestinal flora in AD remains lacking. We induced an AD model through bilateral intracerebroventricular injection of streptozotocin in rats. We divided 36 rats randomly into 6 groups: sham-operated, model, Danggui Shaoyao San (DSS), and 3 DSS decomposed recipes groups. Cognitive abilities were assessed using water maze and open field experiments. Nissl staining examined hippocampal neuron integrity. Western blot analysis determined synaptoprotein expression. Additionally, 16S rDNA high-throughput sequencing analyzed intestinal flora composition. DSS and its decomposed recipe groups demonstrated improved learning and memory in rats (P<0.01). The open field test indicated increased central zone residence time and locomotor activity distance in these groups (P<0.05). Furthermore, the DSS and decomposed recipe groups exhibited reduced hippocampal neuronal damage and increased expression levels of synapsin I (P<0.05) and PSD95 (P<0.01) proteins. Alpha and Beta diversity analyses showed that the intestinal flora species richness and diversity in the DSS and decomposed recipe groups were similar to those in the sham-operated group, signifying a significant restorative effect (P<0.05). The combination of DSS and its decomposed recipes can reduce the abundance of harmful gut microbiota, leading to improvements in cognitive and learning abilities.


[Survey of the existing housing situation for cats (Felis catus) in German-Swiss animal shelters].
This study investigated the current situation of cat husbandry in the animal shelters of german-speaking Switzerland. In 1993, Swiss-german animal shelters occupied just over 5000 housing spaces. Additionally, there was one quarantine space per eight housing spaces. During the summer, which is the season of highest demand, the requirement for housing and quarantine spaces was inadequate. In 1993, approximately 30,000 cats were housed in Swiss-german animal shelters. About four fifths of all entries were boarding cats and the rest were homeless cats. In 1993, about three-quarters of all homeless cats were rehoused with new owners. About a tenth of the remaining cats were reclaimed by their owners. Another tenth of the animals were euthanatized. 63 animal shelters kept cats mainly in group housing enclosures. 15 preferred single/pair boxes, whereas 22 animal shelters used both types of housing. This study shows that more housing and quarantine spaces are needed and should be built for the summer season, to avoid the inadequate housing capacities in Swiss-german animal shelters.


Nutrition and Cardiovascular Health.
Cardiovascular disease (CVD) is the leading cause of death in Western countries, representing almost 30% of all deaths worldwide. Evidence shows the effectiveness of healthy dietary patterns and lifestyles for the prevention of CVD. Furthermore, the rising incidence of CVD over the last 25 years has become a public health priority, especially the prevention of CVD (or cardiovascular events) through lifestyle interventions. Current scientific evidence shows that Western dietary patterns compared to healthier dietary patterns, such as the 'Mediterranean diet' (MeDiet), leads to an excessive production of proinflammatory cytokines associated with a reduced synthesis of anti-inflammatory cytokines. In fact, dietary intervention allows better combination of multiple foods and nutrients. Therefore, a healthy dietary pattern shows a greater magnitude of beneficial effects than the potential effects of a single nutrient supplementation. This review aims to identify potential targets (food patterns, single foods, or individual nutrients) for preventing CVD and quantifies the magnitude of the beneficial effects observed. On the other hand, we analyze the possible mechanisms implicated in this cardioprotective effect.


Radiographic PEEK double-lucency finding after anterior cervical discectomy and fusion with local autograft and PEEK spacer: a preliminary study.
Anterior cervical discectomy and fusion (ACDF) is considered a standard neurosurgical treatment for cervical degenerative disc disease, but the methods for determining fusion after ACDF that involves the use of a polyetheretherketone (PEEK) cage are not well defined. The authors examine an image finding called "PEEK double lucency" that may be useful in identifying fusion. This finding was studied to determine if it would be helpful in identifying fusion on radiographs obtained in 148 patients who underwent an ACDF in which a PEEK cage was filled with local autograft bone (bone spurs in the present cases). The presence of a PEEK double lucency was discovered during review of the plain radiographs and defined as a complete radiolucent ring around the titanium markers within the PEEK cage. In total, 178 spinal levels were examined. Of the 356 (2 for each of the 178 levels) titanium portions examined, the PEEK double lucency was present in 323 or 91% of the titanium portions of the PEEK cages for the patients with complete fusion. The authors conclude that the radiolucent feature of the PEEK cage, which produces the image finding PEEK double lucency, aids in the determination of fusion.


Clinical significance of contact lens related changes of ocular surface tissue observed on optical coherence images.
To investigate the relationship between the real contact lens imprint into the conjunctival tissue, observed by optical coherence tomography (OCT) and conjunctival staining and contact lens wearing comfort. 17 participants (mean age = 26.6 SD ± 3.6 years; 7 females) were fitted with three different contact lenses base curves of the same silicone hydrogel custom lens type (Visell 50; Hecht Contactlinsen, Au, Germany) in a randomised order. One lens was optimally fitted according to the manufacturer's recommendation, one fitted 0.4 mm flatter and one fitted 0.4 mm steeper. After 4 h of lens wear the contact lens edge in the area of the conjunctiva was imaged nasally and temporally using OCT (Optovue iVue SD-OCT). To correct the artefact due to optical distortion with OCT, the imprint of all worn lenses was measured on a glass plate afterwards. Conjunctival staining in the limbal region after 4 h of lens wear was classified using the CCLRU Grading Scale. Comfort scoring was based on visual analog scales from 0 (very poor) to 100 (excellent). The mean conjunctival imprint of all contact lens edges was 32.0 ± 8.1 μm before and 7.3 ± 6.5 μm after distortion correction of the OCT images. The distortion corrected conjunctival imprint with the 0.4 mm steeper lens (11.5 ± 6.2 μm) was statistically significantly greater compared to the optimally fitted lens (6.5 ± 5.9 μm) (One-way ANOVA followed Tukey-test; p = 0.017) and greater compared to the 0.4 mm flatter lens (3.9 ± 5.3 μm) (p < 0.001). There was no statistically significant difference between the optimally fitted lens and the 0.4 mm flatter lens (p = 0.209). The nasally measured imprint (11.4 ± 9.0 μm) was significantly greater than the temporally measured (3.3 ± 7.6 μm) (p < 0.001). There was no statistically significant correlation between the amount of conjunctival imprint and the graded conjunctival staining (p = 0.346) or the wearer's comfort (p = 0.735). Contact lens edges imaged by OCT exhibited displacement artefacts. The observed conjunctival imprints are a combination of real conjunctival compression and artefacts. A deeper imprint of the contact lens into the conjunctiva caused by a steeper base curve was not related to clinically significant staining or changes in comfort after 4 h of lens wear. The observed differences between nasal and temporal imprint are likely to be caused by variations of conjunctival thickness and the shape of the underlying sclera.


Selection, design and evaluation of new radioligands for PET studies of cardiac adrenoceptors.
Changes in the numbers of human cardiac adrenoceptors (ARs) are associated with various diseases, such as myocardial ischemia, congestive heart failure, cardiomyopathy and hypertension. There is a clear need for capability to assess human cardiac ARs directly in vivo. Positron emission tomography (PET) is an imaging technique that provides this possibility, if effective radioligands can be developed for the targeted ARs. Here, the status of myocardial AR radioligand development for PET is described. Currently, there exist effective radioligands for imaging beta-ARs in human myocardium. One of these, [11C](S)-CGP 12177, is applied extensively to clinical research with PET, sometimes with other tracers of other aspects of the noradrenalin system. Alternative radioligands are in development for beta-ARs, including beta 1-selective radioligands. A promising radioligand for imaging myocardial alpha 1-ARs, [11C]GB67, is now being evaluated in human PET experiments.


Sensitive quantification of α-glucans in mouse tissues, cell cultures, and human cerebrospinal fluid.
The soluble α-polyglucan glycogen is a central metabolite enabling transient glucose storage to suit cellular energy needs. Glycogen storage diseases (GSDs) comprise over 15 entities caused by generalized or tissue-specific defects in enzymes of glycogen metabolism. In several, e.g. in Lafora disease caused by the absence of the glycogen phosphatase laforin or its interacting partner malin, degradation-resistant abnormally structured insoluble glycogen accumulates. Sensitive quantification methods for soluble and insoluble glycogen are critical to research, including therapeutic studies, in such diseases. This paper establishes methodological advancements relevant to glycogen metabolism investigations generally, and GSDs. Introducing a pre-extraction incubation method, we measure degradation-resistant glycogen in as little as 30 mg of skeletal muscle or a single hippocampus from Lafora disease mouse models. The digestion-resistant glycogen correlates with the disease-pathogenic insoluble glycogen and can readily be detected in very young mice where glycogen accumulation has just begun. Second, we establish a high-sensitivity glucose assay with detection of ATP depletion, enabling 1) quantification of α-glucans in cell culture using a medium-throughput assay suitable for assessment of candidate glycogen synthesis inhibitors, and 2) discovery of α-glucan material in healthy human cerebrospinal fluid, establishing a novel methodological platform for biomarker analyses in Lafora disease and other GSDs.


Intra- and Interobserver Reproducibility of Third Trimester Middle Cerebral Artery Pulsatility Index Measurement: A Prospective Cross-Sectional Study.
Middle cerebral artery (MCA) pulsatility index (PI) Doppler in the third trimester of pregnancy is increasingly used. The aim of the study was to investigate intra- and interobserver reproducibility of MCA PI in the third trimester. Singleton pregnancies between 30+0 and 40+0 weeks were recruited. MCA Doppler velocimetry measurements were performed prospectively, independently, and blindly. Intra- and interobserver reproducibility was assessed by concordance correlation coefficient (CCC) and intraclass correlation coefficient (ICC); Bland-Altman plots were built, and limits of agreement (LoA) were calculated. Results were interpreted according to the cutoff set by the True Reproducibility of Ultrasound Techniques Review. We enrolled 101 patients. ICCs for intraobserver reproducibility were 0.84 and 0.78 for raw values and percentiles, respectively; CCCs were 0.72 and 0.64. For interobserver reproducibility ICCs were 0.84 and 0.78, CCCs 0.72 and 0.63. According to the chosen criteria, these values show a poor-moderate reproducibility of third trimester MCA PI. Cohen's Kappa coefficients were 0.59 and 0.42, indicating a moderate agreement in discriminating normal and abnormal values. Intra- and interobserver reproducibility of third trimester MCA PI, as assessed by ICC, CCC, and LoA, is far from satisfactory. This should be taken into account before taking clinical decisions.


Gadobenate dimeglumine-enhanced magnetic resonance imaging of primary leiomyoma of the liver.
We report a case of histologically proven primary leiomyoma of the liver that was evaluated with multiphasic 64-section computed tomography (CT) and gadobenate dimeglumine-enhanced magnetic resonance (MR) imaging. This lesion showed vivid enhancement during the arterial phase with sustained enhancement during the hepatic venous and equilibrium phases. During the liver-specific MR imaging phase (150 minutes after contrast injection), the same lesion demonstrated lack of contrast retention, thus appearing hypointense compared with the background liver. Because of this latter finding, the patient underwent partial resection of the liver. In primary hepatic leiomyoma, the absence of contrast uptake during the liver-specific phase of gadobenate dimeglumine-enhanced MR imaging may be inappropriately interpreted as a sign of malignancy, thus leading to unnecessary, aggressive management of such lesions.


Model-Based ISO 14971 Risk Management of EEG-Based Medical Devices.
Risk management (RM) is a key component of the development of modern medical devices (MD) to achieve acceptable functional safety and pass the regulatory process. The emerging availability of various techniques, languages, and tools that use model-based system engineering (MBSE) promises to facilitate the development and analysis of complex MD. In this paper, we show how to integrate RM principles and activities recommended in ISO 14971 medical standard into an MBSE-driven MD development process. We propose a method and framework capable of modeling essential RM concepts and performing RM and safety analysis in the early stages of the MD development life cycle. The framework extends OMG RAAML (Object Management Group Risk Analysis and Assessment Modeling Language) to the medical domain according to ISO 14971. We illustrate our approach using a case study of the e-Glass system developed for real-time EEG-based subject monitoring with the intended use of stress monitoring.Clinical Relevance-This facilitates the MD certification process by semi-automation of RM based on ISO 14971 and obtaining safe MD by design.


Morphological redescription of Dirofilaria immitis.
Morphological descriptions of Dirofilaria immitis are scarce. For this reason, we carried out morphological studies using both light and scanning electron microscopy for this filaroid species. Morphometric and morphological data were compatible with previous descriptions of D. immitis , but several anatomical structures are described by scanning electron microscopy for the first time, such as details of the cuticular striations, positioning of amphids, visualization of anal and vulvar opening, descriptions of deirids, lateral line, the pair of phasmids in the posterior end in females, and visualization of a small pair of latero-terminal papillae in the posterior end in males.


Impact of adjuvant therapy in patients with invasive intraductal papillary mucinous neoplasms of the pancreas: an international multicenter cohort study.
Adjuvant therapy prolongs survival in patients with pancreatic ductal adenocarcinoma. However, no clear guidelines are available regarding the oncologic effects of adjuvant therapy (AT) in resected invasive intraductal papillary mucinous neoplasms (IPMN). The aim was to investigate the potential role of AT in patients with resected invasive IPMN. From 2001 to 2020, 332 patients with invasive pancreatic IPMN were retrospectively reviewed in 15 centres in eight countries. Propensity score-matched and stage-matched survival analyses were conducted. A total of 289 patients were enroled in the study after exclusion (neoadjuvant therapy, unresectable disease, uncertain AT status, and stage IV). A total of 170 patients were enroled in a 1:1 propensity score-matched analysis according to the covariates. In the overall cohort, disease-free survival was significantly better in the surgery alone group than in the AT group ( P =0.003), but overall survival (OS) was not ( P =0.579). There were no significant differences in OS in the stage-matched analysis between the surgery alone and AT groups (stage I, P =0.402; stage II, P =0.179). AT did not show a survival benefit in the subgroup analysis according to nodal metastasis (N0, P =0.481; N+, P =0.705). In multivariate analysis, node metastasis (hazard ratio, 4.083; 95% CI, 2.408-6.772, P <0.001), and cancer antigen 19-9 greater than or equal to 100 (hazard ratio, 2.058; 95% CI, 1.247-3.395, P =0.005) were identified as adverse prognostic factors in resected invasive IPMN. The current AT strategy may not be recommended to be performed with resected invasive IPMN in stage I and II groups, unlike pancreatic ductal adenocarcinoma. Further investigations of the potential role of AT in invasive IPMN are recommended.


Purification, characterization and amino-acid sequence analysis of a thermostable, low molecular mass endo-beta-1,4-glucanase from blue mussel, Mytilus edulis.
A cellulase (endo-beta-1,4-D-glucanase, EC 18.104.22.168) from blue mussel (Mytilus edulis) was purified to homogeneity using a combination of acid precipitation, heat precipitation, immobilized metal ion affinity chromatography, size-exclusion chromatography and ion-exchange chromatography. Purity was analyzed by SDS/PAGE, IEF and RP-HPLC. The cellulase (endoglucanase) was characterized with regard to enzymatic properties, isoelectric point, molecular mass and amino-acid sequence. It is a single polypeptide chain of 181 amino acids cross-linked with six disulfide bridges. Its molecular mass, as measured by MALDI-MS, is 19 702 Da; a value of 19 710.57 Da was calculated from amino-acid composition. The isoelectric point of the enzyme was estimated by isoelectric focusing in a polyacrylamide gel to a value of 7.6. According to amino-acid composition, the theoretical pI is 7.011. The effect of temperature on the endoglucanase activity, with carboxymethyl cellulose and amorphous cellulose as substrates, respectively, was studied at pH 5.5 and displayed an unusually broad optimum activity temperature range between 30 and 50 degrees C. Another unusual feature is that the enzyme retains 55-60% of its maximum activity at 0 degrees C. The enzyme readily degrades amorphous cellulose and carboxymethyl cellulose but displays no hydrolytic activity towards crystalline cellulose (Avicel) and shows no cross-specificity for xylan; there is no binding to Avicel. The enzyme can withstand 10 min at 100 degrees C without irreversible loss of enzymatic activity. Amino-acid sequence-based classification has revealed that the enzyme belongs to the glycoside hydrolase family 45, subfamily 2 (B. Henrissat, Centre de Recherches sur les Macromolecules Végétales, CNRS, Joseph Fourier Université, Grenoble, France, personal communication).


Colorectal specialization and survival in colorectal cancer.
It is recognized that higher surgeon volume is associated with improved survival in colorectal cancer. However, there is a paucity of national studies that have evaluated the relationship between surgical specialization and survival. We used the Surveillance, Epidemiology, and End Results Medicare cancer registry to examine the association between colorectal specialization (CRS) and disease-specific survival (DSS) between 2001 and 2009. A total of 21,432 colon cancer and 5893 rectal cancer patients who underwent elective surgical resection between 2001 and 2009 were evaluated. Univariate and multivariate Cox survival analysis was used to identify the association between surgical specialization and cancer-specific survival. Colorectal specialists performed 16.3% of the colon and 27% of the rectal resections. On univariate analysis, specialization was associated with improved survival in Stage II and Stage III colon cancer and Stage II rectal cancer. In multivariate analysis, however, CRS was associated with significantly improved DSS only in Stage II rectal cancer [hazard ratio (HR) 0.70, P = 0.03]. CRS was not significantly associated with DSS in either Stage I (colon HR 1.14, P = 0.39; rectal HR 0.1.26, P = 0.23) or Stage III (colon HR 1.06, P = 0.52; rectal HR 1.08, P = 0.55) disease. When analysis was limited to high volume surgeons only, the relationship between CRS and DSS was unchanged. CRS is associated with improved DSS following resection of Stage II rectal cancer. A combination of factors may contribute to long-term survival in these patients, including appropriate surgical technique, multidisciplinary treatment decisions and guideline-adherent surveillance. CRS probably contributes positively to these factors resulting in improved survival.


Comparative Healthcare Resource Utilization of Percutaneous Mechanical Circulatory Support Using Impella Versus Intra-aortic Balloon Pump Use for Patients With Acute Coronary Syndrome and Cardiogenic Shock Undergoing Percutaneous Coronary Interventions: Insights From National Inpatient Sample.
The use of percutaneous mechanical circulatory support (MCS) devices, including Impella and Intra-aortic balloon pump (IABP), in patients with cardiogenic shock has increased in recent times. We aimed to evaluate the impact of the choice of an MCS device on healthcare resource utilization. We queried the National Inpatient Sample registry between October 2016 and December 2018 to identify adults admitted for acute coronary syndrome-related cardiogenic shock and who received percutaneous coronary intervention (PCI). The study population was segregated into Impella and IABP groups using ICD 10 diagnosis codes. The primary endpoint was high healthcare resource utilization (HRU), while secondary outcomes included periprocedural complications. Propensity scoring matching was used to determine which patients in the Impella cohort had similar health to IABP patients. During the study period, 439,610 patients were admitted who received hemodynamic support using, Impella or IABP on account of acute coronary syndrome complicated by cardiogenic shock (CS). The median age (years) of the Impella cohort and IABP cohorts were similar (64.1 vs 65.1, P = 0.08). Gender distribution of the Impella CS patients was like IABP patients with female majorities in both groups, (71.9% vs 67.9%, P = 0.05). Impella CS patients had a higher representation of those with hypertension (P = 0.002), smoking (P = 0.040), obesity (P = 0.034), diabetes mellitus (P = 0.009), CHF (P = 0.030), COPD (P = 0.034), chronic liver disease (P = 0.028), and chronic kidney disease (P = 0.031). 1:1 Propensity score matching identified 2620 Impella patients' comparable severity index with the IABP patients. Patients with hemodynamic support using Impella had higher healthcare resource utilization, (HRU), the surrogate of length of stay (LOS) ≥7 or nonhome disposition at discharge, when compared with those with IABP (57.41% vs 42.76%, P < 0.0001). Impella CS patients had higher in-hospital mortality as compared to the IABP patients (55.45% vs 45.86%, P < 0.0001). Impella CS patients developed more periprocedural complications, including vascular injury (4.8% vs 1.4%, P < 0.0001), acute kidney injury (58.36% vs 41.64%, P < 0.0001), end-stage renal disease requiring dialysis (8.75% vs 1.25%, P = 0.002) when compared to the IABP patients. Among patients with ACS undergoing PCI and receiving MCS devices, those receiving Impella demonstrated higher healthcare resource utilization, higher LOS ≥7 days, and more nonhome disposition at discharge compared to patients receiving IABP. Further investigation is warranted to elucidate factors associated with these findings.


Longitudinal progression of movement abnormalities in relation to psychotic symptoms in adolescents at high risk of schizophrenia.
Because recent findings suggest that early treatment may ameliorate the course or even prevent the onset of schizophrenia and other psychotic disorders, longitudinal high-risk research on biological markers of risk has become a priority. Within this context, premorbid movement abnormalities are of particular interest because the neurocircuitry hypothesized to give rise to dyskinetic movements has also been implicated in psychotic symptoms. To date, there have been no published longitudinal studies examining the progression of movement abnormalities and their relation with symptom progression in at-risk youth. To examine the progression of movement abnormalities in relation to positive and negative symptoms in adolescents at high risk of developing psychotic disorders. Naturalistic, prospective, longitudinal design. Participants recruited through announcements directed at parents of adolescents showing schizotypal symptoms. One hundred twenty-one adolescents (mean baseline age, 14.26 years), 32 with schizotypal personality disorder, 49 nonclinical controls, and 40 with other personality disorders. Participating adolescents were evaluated for personality disorders (Structured Interview for DSM-IV Personality Disorders), prodromal symptoms (Structured Interview for Prodromal Symptoms), and movement abnormalities (Dyskinesia Identification System Condensed User Scale) at 3 annual assessments. The schizotypal group exhibited significantly elevated movement abnormalities in comparison with controls across all 3 time points. Further, the schizotypal personality disorder group alone showed significant increases in movement abnormalities over time. Movement abnormalities were correlated with prodromal symptoms at each time period, and for several body regions, the magnitude of this relationship significantly increased over time. The results are consistent with the hypothesis of shared neural circuitry for movement abnormalities and psychotic symptoms and suggest the potential value of including an assessment of motor signs in screening for psychosis risk.


Corticocortical efferent systems in the monkey: a quantitative spatial analysis of the tangential distribution of cells of origin.
The laminar and tangential distributions of association neurons projecting from areas 4 and 6 of the frontal lobe to area 5 of the superior parietal lobule were studied in macaque monkeys by using horseradish peroxidase histochemistry. In both areas 4 and 6 association neurons were medium-large pyramidal cells of layers II and III, and pyramidal and fusiform cells of layers V-VI. Tangentially, they were distributed unevenly over the cortical surface occupying only certain parts of areas 4 and 6, including the dorsomedial part of area 6, the proximal arm region of Woolsey's M1 map, parts of the postarcuate cortex, and the supplementary motor area. Within these projection zones, the number of projection cells waxed and waned in a periodic fashion. Quantitative methods, including spectral analysis techniques, were used to characterize precisely spatial periodicities along the rostrocaudal dimension. The same quantitative analyses were used to determine the nature of the tangential distribution of corticocallosal cells of area 5 projecting to contralateral area 5. Both association and callosal spectra contained a strong component in the range of low spatial frequencies, corresponding to periods greater than 2 mm. Moreover, a consistent peak was observed in both spectra at spatial frequencies corresponding to periods ranging from 0.85 to 1.28 mm. This peak is in accord with the hypothesis of a modular organization of the cells of origin of these projection systems.


A novel cell nuclei segmentation method for 3D C. elegans embryonic time-lapse images.
Recently a series of algorithms have been developed, providing automatic tools for tracing C. elegans embryonic cell lineage. In these algorithms, 3D images collected from a confocal laser scanning microscope were processed, the output of which is cell lineage with cell division history and cell positions with time. However, current image segmentation algorithms suffer from high error rate especially after 350-cell stage because of low signal-noise ratio as well as low resolution along the Z axis (0.5-1 microns). As a result, correction of the errors becomes a huge burden. These errors are mainly produced in the segmentation of nuclei. Thus development of a more accurate image segmentation algorithm will alleviate the hurdle for automated analysis of cell lineage. This paper presents a new type of nuclei segmentation method embracing an bi-directional prediction procedure, which can greatly reduce the number of false negative errors, the most common errors in the previous segmentation. In this method, we first use a 2D region growing technique together with the level-set method to generate accurate 2D slices. Then a modified gradient method instead of the existing 3D local maximum method is adopted to detect all the 2D slices located in the nuclei center, each of which corresponds to one nucleus. Finally, the bi-directional prediction method based on the images before and after the current time point is introduced into the system to predict the nuclei in low quality parts of the images. The result of our method shows a notable improvement in the accuracy rate. For each nucleus, its precise location, volume and gene expression value (gray value) is also obtained, all of which will be useful in further downstream analyses. The result of this research demonstrates the advantages of the bi-directional prediction method in the nuclei segmentation over that of StarryNite/MatLab StarryNite. Several other modifications adopted in our nuclei segmentation system are also discussed.


Structural characteristics of halobacterial gas vesicles.
Gas vesicle formation in halophilic archaea is encoded by a DNA region (the vac region) containing 14 different genes: gvpACNO and gvpDEFGHIJKLM. In Halobacterium salinarum PHH1 (which expresses the p-vac region from plasmid pHH1), gas vesicles are spindle shaped, whereas predominantly cylindrical gas vesicles are synthesized by the chromosomal c-vac region of H. salinarum PHH4 and the single chromosomal mc-vac region of Haloferax mediterranei. Homologous complementation of gvp gene clusters derived from the chromosomal c-vac region led to cylindrical gas vesicles in transformants and proved that the activity of the c-gvpA promoter depended on a gene product from the c-gvpE-M DNA region. Heterologous complementation experiments with transcription units of different vac regions demonstrated that the formation of chimeric gas vesicles was possible. Comparison of micrographs of wild-type and chimeric gas vesicles indicated that the shape was not exclusively determined by GvpA, the major structural protein of the gas vesicle wall. More likely, a dynamic equilibrium of several gvp gene products was responsible for determination of the shape. Transmission electron microscopy of frozen hydrated, wild-type gas vesicles showed moiré patterns due to the superposition of the front and back parts of the ribbed gas vesicle envelope. Comparison of projections of model helices with the moiré pattern seen on the cylindrical part of the gas vesicles provided evidence that the ribs formed a helix of low pitch and not a stack of hoops.


Cell specificity of transcription regulation by papovavirus T antigens and DNA replication.
Simian virus 40 (SV40) and polyomavirus (Py) DNA replication require cellular proteins and a virus-encoded early gene product, large T antigen (SVT and PyT, respectively). Primate cells contain factors permissive for SV40 replication, whereas murine cells express those factors permissive for Py. We have compared the roles T antigen, cell permissiveness and replication play in transcription of SV40 and Py genes. We show that in their respectively permissive cells, SV40 replication causes a major shift in transcription initiation from the early to the late viral promoter, whereas when Py replicates a comparable shift does not occur. This difference is discussed in relation to differences in the organization of the origin and promoter region between these two papovaviruses. Reporter plasmids were constructed that carried both viral origins, one at the natural position in the promoter being tested and the other at a distal location. With the appropriate TAg, these vectors could be made to replicate in either primate (HeLa) or rodent (3T6) cells. The SV40 early to late shift occurred when replication was driven in HeLa cells, and was not seen on replicating templates in rodent cells. Thus, replication per se does not account for the shift. We show also that, like SVT, PyT is a potent activator of transcription, and that SVT and PyT can activate each other's late promoters independently of DNA replication, but only in cells permissive for DNA replication catalysed by the respective T antigen. Taken together, the data presented here suggest that papovaviruses may utilize permissive factors in transcription control mechanisms.


Anti-tumour effects of antimicrobial peptides, components of the innate immune system, against haematopoietic tumours in Drosophila mxc mutants.
The innate immune response is the first line of defence against microbial infections. In Drosophila, two major pathways of the innate immune system (the Toll- and Imd-mediated pathways) induce the synthesis of antimicrobial peptides (AMPs) within the fat body. Recently, it has been reported that certain cationic AMPs exhibit selective cytotoxicity against human cancer cells; however, little is known about their anti-tumour effects. Drosophila mxcmbn1 mutants exhibit malignant hyperplasia in a larval haematopoietic organ called the lymph gland (LG). Here, using RNA-seq analysis, we found many immunoresponsive genes, including those encoding AMPs, to be upregulated in these mutants. Downregulation of these pathways by either a Toll or imd mutation enhanced the tumour phenotype of the mxc mutants. Conversely, ectopic expression of each of five different AMPs in the fat body significantly suppressed the LG hyperplasia phenotype in the mutants. Thus, we propose that the Drosophila innate immune system can suppress the progression of haematopoietic tumours by inducing AMP gene expression. Overexpression of any one of the five AMPs studied resulted in enhanced apoptosis in mutant LGs, whereas no apoptotic signals were detected in controls. We observed that two AMPs, Drosomycin and Defensin, were taken up by circulating haemocyte-like cells, which were associated with the LG regions and showed reduced cell-to-cell adhesion in the mutants. By contrast, the AMP Diptericin was directly localised at the tumour site without intermediating haemocytes. These results suggest that AMPs have a specific cytotoxic effect that enhances apoptosis exclusively in the tumour cells.


Postnatal regulation of 2,3-DPG in sheep erythrocytes.
The erythrocyte 2,3-diphosphoglycerate (2,3-DPG) concentrations of sheep change markedly during the 1st mo following birth. From measurements of erythrocyte glycolytic enzymes and intermediate concentrations, we have identified the mechanism regulating erythrocyte 2,3-DPG in postnatal sheep. The postnatal changes in erythrocyte 2,3-DPG do not result from qualitative or quantitative changes in the intracellular activities of the Rapoport-Luebering shunt enzymes, 2,3-DPG mutase or 2,3-DPG phosphatase. The postnatal 2,3-DPG changes result from changes in the erythrocyte concentration of 1,3-DPG, which is controlled by other reactions in the glycolytic pathway. Neither changes in the glycolytic control enzymes (hexokinase, phosphofructokinase, and pyruvate kinase) nor changes in the intrinsic glycolytic rate can account for these 1,3-DPG concentration changes. 1,3-DPG concentrations are regulated by the in vivo glycolytic rate, which is controlled by the intracellular concentration of glucose, the glycolytic substrate. Glucose concentrations are 0.3 mmol/l cells in erythrocytes of fetal sheep (135-140 days gestational age), increase following birth to a peak of 3.8 mmol/l cells by the 1st wk of age, and then decline to the normal adult levels of 0.5 mmol/l cells by the end of the 1st mo.


[In vivo study of the pharmacokinetic interaction of afobazole and losartan (cytochrome CYP2C9 substrate)].
The influence of afobazole on isoenzyme CYP2C9 production in rats was studied using losartan as the marker drug. Single dose of losartan was administered orally without afobazole in a dose of 30 mg/kg and in the same single (30 mg/kg) on the background of 3- and 4-day administration of afobazole in a dose of 5, 25, 75, 100, and 125 mg/kg. At 5 mg/kg (effective dose for anxiolytic effect), afobazole did not cause any induction/inhibition effect on CYP2C9 isoenzyme. A multiple increase in afobazole dose was manifested by a moderate induction effect. The maximum induction effect of afobazole was achieved in a dose of 75 mg/kg. At doses above 75 mg/kg, the induction effect of afobazole was less pronounced.


Drug repurposing for Alzheimer's disease from 2012-2022-a 10-year literature review.
Background: Alzheimer's disease (AD) is a debilitating neurodegenerative condition with few treatment options available. Drug repurposing studies have sought to identify existing drugs that could be repositioned to treat AD; however, the effectiveness of drug repurposing for AD remains unclear. This review systematically analyzes the progress made in drug repurposing for AD throughout the last decade, summarizing the suggested drug candidates and analyzing changes in the repurposing strategies used over time. We also examine the different types of data that have been leveraged to validate suggested drug repurposing candidates for AD, which to our knowledge has not been previous investigated, although this information may be especially useful in appraising the potential of suggested drug repurposing candidates. We ultimately hope to gain insight into the suggested drugs representing the most promising repurposing candidates for AD. Methods: We queried the PubMed database for AD drug repurposing studies published between 2012 and 2022. 124 articles were reviewed. We used RxNorm to standardize drug names across the reviewed studies, map drugs to their constituent ingredients, and identify prescribable drugs. We used the Anatomical Therapeutic Chemical (ATC) Classification System to group drugs. Results: 573 unique drugs were proposed for repurposing in AD over the last 10 years. These suggested repurposing candidates included drugs acting on the nervous system (17%), antineoplastic and immunomodulating agents (16%), and drugs acting on the cardiovascular system (12%). Clozapine, a second-generation antipsychotic medication, was the most frequently suggested repurposing candidate (N = 6). 61% (76/124) of the reviewed studies performed a validation, yet only 4% (5/124) used real-world data for validation. Conclusion: A large number of potential drug repurposing candidates for AD has accumulated over the last decade. However, among these drugs, no single drug has emerged as the top candidate, making it difficult to establish research priorities. Validation of drug repurposing hypotheses is inconsistently performed, and real-world data has been critically underutilized for validation. Given the urgent need for new AD therapies, the utility of real-world data in accelerating identification of high-priority candidates for AD repurposing warrants further investigation.


Chevron formation of the zebrafish muscle segments.
The muscle segments of fish have a folded shape, termed a chevron, which is thought to be optimal for the undulating body movements of swimming. However, the mechanism shaping the chevron during embryogenesis is not understood. Here, we used time-lapse microscopy of developing zebrafish embryos spanning the entire somitogenesis period to quantify the dynamics of chevron shape development. By comparing such time courses with the start of movements in wildtype zebrafish and analysing immobile mutants, we show that the previously implicated body movements do not play a role in chevron formation. Further, the monotonic increase of chevron angle along the anteroposterior axis revealed by our data constrains or rules out possible contributions by previously proposed mechanisms. In particular, we found that muscle pioneers are not required for chevron formation. We put forward a tension-and-resistance mechanism involving interactions between intra-segmental tension and segment boundaries. To evaluate this mechanism, we derived and analysed a mechanical model of a chain of contractile and resisting elements. The predictions of this model were verified by comparison with experimental data. Altogether, our results support the notion that a simple physical mechanism suffices to self-organize the observed spatiotemporal pattern in chevron formation.


Maternal occupation in agriculture and risk of limb defects in Washington State, 1980-1993.
This study examined the association between maternal occupational exposure to agricultural chemicals and the risk of limb defects among offspring. A retrospective cohort study was conducted using Washington State birth records for the years 1980 through 1993. The exposed group, consisting of 4466 births to mothers employed in agriculture, was compared with 2 reference groups: (i) 23,512 births in which neither parent worked in agriculture ("nonagricultural" group) and (ii) 5994 births in which only the father worked in agriculture ("paternal agriculture" group). The outcome of interest was limb defects [syndactyly, polydactyly, adactyly, and "other limb reductions" (as listed in the birth record)]. An elevated risk of limb defects was observed for the exposed group in comparison with both the nonagricultural and paternal agriculture groups, with ethnicity-adjusted prevalence ratios of 2.6 [95% confidence interval (95% CI) 1.1-5.8] and 2.6 (95% CI 0.7-9.5), respectively. These results support the hypothesis that maternal occupational exposure to agricultural chemicals may increase the risk of giving birth to a child with limb defects.


CO2 wetting on pillar-nanostructured substrates.
CO2 capture by dropwise CO2 condensation on cold solid surfaces is a promising technology. Understanding the role of the nanoscale surface and topographical features of CO2 droplet wetting characteristics is of importance for CO2 capture by this technology, but this remains unexplored as of yet. Here, using large-scale molecular dynamics (MD) simulations, the contact angle and wetting behaviors of CO2 droplets on pillar-structured Cu-like surfaces are investigated for the first time. Dynamic wetting simulations show that, by changing the strength of the solid-liquid attraction [Formula: see text] a smooth Cu-like surface offers a transition from CO2-philic to CO2-phobic. By periodically pillared roughening of the Cu-like surfaces, however, a higher contact angle and a smaller spreading exponent of a liquid CO2 droplet are realized. Particularly, a critical crossover of CO2-philic to CO2-phobic can appear. The wetting of the pillared surfaces by a liquid CO2 droplet proceeds non-uniformly. A liquid CO2 droplet is capable of exhibiting a transition from the Cassie state to the Wenzel state with increasing [Formula: see text] increasing inter-pillar distance, and increasing pillar width. The wetting morphologies of the metastable Wenzel state of a CO2 droplet are very different from each other. The findings will inform the ongoing design of CO2-phobic solid surfaces for practical dropwise condensation-based CO2 capture applications.


Prenatal exposure to phthalates and neuropsychological development during childhood.
There is inconsistent evidence regarding the effects of prenatal phthalate exposure on children's neuropsychological development. We evaluate the association between prenatal phthalate exposure and the cognitive, psychomotor and behavioral development of 367 children at repeated ages in a prospective birth cohort study. We measured phthalate metabolites (sum of four DEHP metabolites - Σ4DEHP, MBzP, MEP, MiBP and MnBP) in urine samples collected during the 1st and 3rd trimesters of pregnancy in women participating in the INMA-Sabadell birth cohort study. We assessed cognitive and psychomotor development of their children at 1 and 4 years, and social competence, ADHD symptoms and other behavioral problems at 4 and 7 years. No associations were observed between prenatal phthalate exposure and cognitive and psychomotor scores at the age of 1 year and at the age of 4 years, except for an association between MBzP and lower psychomotor scores (β=-1.49 [95% confidence interval (CI)=-2.78, -0.21]). Σ4DEHP concentrations were associated with increased social competence scores at 4 years and with reduced ADHD symptoms at age 4 and 7 years. Increasing MEP concentrations were associated with a reduced risk of inattention symptoms at 4 years. No associations were observed for MBzP, MiBP or MnBP in relation to behavioral problems. This study, with multiple phthalate exposure measurements and measures of neuropsychological domains at different ages, suggest that prenatal phthalate exposure does not adversely affect children's cognitive, psychomotor or behavioral development.


Genetical and molecular analyses of qa-2 transformants in Neurospora crassa.
Neurospora crassa qa-2+ transformants from five different donor DNA clones were analyzed by genetical and molecular techniques. None of the 32 transformants have the qa-2+ DNA replacing the qa-2- gene in linkage group VII. In one transformant, the qa-2+ DNA was inserted adjacent to the qa-2- gene. Thirty-one transformants have the qa-2+ inserts at sites not linked, or not closely linked, to the qa-2 locus in LG VII. Plasmid sequences were integrated along with the qa-2+ gene in 28 transformants. In the unlinked duplication-type transformants, catabolic dehydroquinase (the qa-2+ gene product) was induced at 5-100% of the wild-type-induced enzyme activity, with 24 transformants in the 5-80% range. The reduced levels of enzyme activity may be due to "position effects" of sequences adjacent to the integration site either in the N. crassa genomic DNA or in the flanking plasmid (pBR322 or pBR325) sequences. Unexpected gene conversion-like events, in which a qa-2+ gene was changed to qa-2-, were observed in tetrads from intercrosses between unlinked duplication-type transformants and in selfings of such transformants.